A rapid mix flow cytometer with subsecond kinetic resolution

Cytometry
J P NolanL A Sklar

Abstract

Kinetic approaches are valuable tools for mechanistic studies of cell function. Flow cytometry is well suited to make sensitive kinetic measurements, but the time required to deliver mixed samples to the point of measurement (10-20 s in a conventional cytometer) limits analysis of rapidly occurring events. To address this limitation, we adapted a syringe-based stopped-flow rapid mixing device to a modified commercial flow cytometer to achieve mixing and measurement of sample in under 1 s. Because such screw-driven mixers are designed to deliver fluid at rates of microliters per millisecond and cytometers accept samples at microliters per second, the syringe mixer was modified with a screw to allow sample delivery at rates as low as 1.8 microliters/s. A custom-made nozzle holder featuring a fast-acting three-way sample delivery valve and a 1.5- microliters dead volume was designed for a Becton Dickinson FACS stream-in-air flow nozzle. Syringe motors and valves are computer controlled, as is the start signal for an adjustable time ramp. A stable sample stream can be established within the sheath stream in less than 1 s, enabling fluorescence measurements of microspheres with coefficients of variation of approximately 5%. Light sc...Continue Reading

References

Jan 1, 1987·Annual Review of Biophysics and Biophysical Chemistry·L A Sklar
Sep 1, 1985·Cytometry·J H Jett, R G Alexander
Jan 11, 1980·Science·J C Martin, D E Swartzendruber
Mar 1, 1981·Cytometry·R D HiebertG C Salzman

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Citations

Dec 21, 2000·Cytometry·S W GravesJ P Nolan
Sep 10, 2002·Molecular Immunology·Richard G PosnerWilliam S Hlavacek
Dec 18, 2001·Trends in Biotechnology·John P Nolan, Larry A Sklar
May 14, 1999·Drug Discovery Today·J P NolanL A Sklar
Feb 4, 2014·Lab on a Chip·Menake E Piyasena, Steven W Graves
Dec 31, 2002·Journal of Receptor and Signal Transduction Research·John C WilkinsonJames V Staros
Aug 6, 2005·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Yang WuTione Buranda
Aug 8, 2007·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Mark A NaivarJames P Freyer
Nov 3, 2011·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Attila Tárnok
Sep 24, 2013·Biochimica Et Biophysica Acta·Rajashri SridharanMark E Dumont
May 4, 2002·Annual Review of Biophysics and Biomolecular Structure·Larry A SklarEric R Prossnitz
Jul 8, 2003·The Journal of Biological Chemistry·Alexandre ChigaevLarry A Sklar
Jul 10, 2013·Current Protocols in Cytometry·Pearlson P Austin Suthanthiraraj, Steven W Graves

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